华蟾素抑制结肠癌细胞增殖转移的实验研究

目的:观察华蟾素对结肠癌细胞HCT116细胞增殖、迁移的影响,探讨其可能的作用机制。方法:常规体外培养结肠癌细胞HCT116细胞系,采用CCK-8法检测不同浓度华蟾素作用不同时间对HCT116细胞增殖的抑制作用,细胞划痕实验和Transwell小室法检测华蟾素对细胞迁移能力的影响,蛋白质印迹法(Western blot)检测细胞基质金属蛋白酶(MMP)-9、MMP-2、钙黏蛋白(E-Cadherin)的表达水平。结果:CCK-8法检测结果表明,华蟾素对HCT116细胞的生长有明显抑制作用,其抑制作用随药物浓度和作用时间的增加而增强(P0.05);细胞划痕实验和Transwell小室法结果显示,随华蟾素药物浓度的增加,HCT116细胞迁移能力下降(P0.05);Western blot结果表明,华蟾素能够明显抑制HCT116细胞MMP-9的蛋白表达并上调E-Cadherin的蛋白表达(P0.01),但对MMP-2蛋白的表达无显著影响(P0.05)。结论:华蟾素能够抑制人结肠癌细胞的增殖和转移,其机制可能与其抑制MMP-9和上调E-Cadherin的蛋白表达有关。

Cinobufacini inhibits the proliferation and metastasis of HCT116 colon cancer cells

  Objective:To investigate the effects of cinobufacini on the proliferation and metastasis of human colon cancer cell line HCT116 and discuss its possible mechanism.   Methods:The colon cancer cell line HCT116 was cultured in vitro. The inhibitory effects of cinobufacini with different concentrations on the proliferation of HCT116 cells at different timepoints were detected by CCK-8 assay. The effects of cinobufacini on the migration were detected by cell scratch test and Transwell chamber assay. The expressions of MMP-9,MMP-2 and E-Cadherin were detected by Western blot.   Results:The results of CCK-8 assay showed that cinobufacini significantly inhibited the growth of HCT116 cells and the inhibitory effect was enhanced with the increase of drug concentration and time(P<0.05). The results of cell scratch test and Transwell chamber assay showed that with the increase of cinobufotalin concentration,HCT116 cell migration ability was decreased(P<0.05). The results of Western blot showed that cinobufotalin could obviously inhibit the protein expression of MMP-9 and up-regulate the protein expression of E-Cadherin in HCT116 cells(P<0.01),but there was no significant effect on the protein expression of MMP-2(P>0.05).   Conclusion:Cinobufacini can inhibit the proliferation and metastasis of human colon cancer cells. Its mechanism may be related to the inhibition on the protein expression of MMP-9 and up-regulation of E-Cadherin.